An overview of experimental strategies and applications of Accelerated Stability Assessment Program (ASAP) studies of oligonucleotide drug substances and drug products will be presented. Practical considerations related to experimental design and execution specific to oligonucleotides will be presented, along with examples of how ASAP studies may be utilized throughout the development lifecycle of oligonucleotide therapeutics. This includes studies aimed at demonstration of material comparability and to augment platform stability data.

Avidity Biosciences is a clinical stage company developing a new type of modality, Antibody Oligonucleotide Conjugates (AOCs). These therapeutics consist of a monoclonal antibody as well as an oligonucleotide,and are therefore hybrids of a biologic and a small molecule. In this talk, we will discuss workflows to overcome the challenges associated with analytical method development and characterization of these novel hybrid therapeutics.

The quality attributes of siRNA drug substances are affected by used raw materials including customized starting materials such as linkers and conjugates, as well as by the manufacturing process, process parameters, and potential degradation products of the compounds. It is important to increase the understanding of processes and products to allow for improvements of processes and product quality as well as to reduce costs.

Traditional phosphorothioate (PS) oligonucleotide synthesis methods generate stereorandom mixtures comprising up to several hundred thousand molecules, each with distinct stereochemistry. Wave Life Sciences is pioneering the development and manufacture of oligonucleotides with control over the backbone chirality. We will outline the various analytical tools we have deployed to characterize the platform processes that generate stereopure oligonucleotides, and unambiguously determine the structural composition, identity, and stability of stereopure oligonucleotides.

Effective change management is an essential part of drug product lifecycle. Typical changes include pre- and post-approval changes to drug substance and drug product manufacturing processes and analytical procedures and supply of raw materials to address regulatory, safety, quality, and supply risks. This presentation will share a phase-appropriate, risk-based approach to understand the risk on critical quality attributes and demonstration of comparability.

With limited regulatory guidance on the acceptable levels of impurities in oligonucleotides, ambiguity regarding specifications and comparability assessment expectations exists. While impurities are anticipated to exhibit the same safety profile as the parent oligonucleotide, they likely offer no inherent benefit and are expected to be controlled. This work outlines an approach to bridge process capability and oligonucleotide impurity safety from a toxicology and immunogenicity perspective to support specifications and comparability.

Double stranded oligonucleotides with multiple 2’substituents, and several phosphorothioate linkages lead to significant numbers of related impurities. Denaturing UPLC methods to separate single strands and their impurities yields complex and often poorly resolved peaks. Non-denaturing methods are often not specific for duplexed impurities. Despite these challenges, sponsors are required to produce a control strategy that identifies and quantifies impurities. In this presentation we will discuss the challenges, strategies to overcome them.

The complexity of oligonucleotide drug candidates being selected for clinical development has increased in recent years. These oligonucleotides often have significant chemical modifications requiring novel starting materials as well as technical innovations in process development, analytical chemistry, manufacturing and controls. This places extraordinary demands on both sponsor companies and contract manufacturing organizations to meet regulatory expectations under aggressive timelines.

For complex drug substances such as oligonucleotides, it is helpful, and sometimes even necessary, to introduce impurity controls throughout the process and not just on the final specification. This presentation will explore opportunities to leverage ICH guidelines to create a comprehensive control strategy, including examples of successes and failures, in a variety of areas, such as starting materials, elemental impurities, and the use of orthogonal HPLC methods.

This talk will focus on the FDA Assessment Aid, a voluntary submission to facilitate FDA assessment of NDA/BLA applications in the Office of Oncologic Drugs. The talk will go over important considerations to include in the pharmacology and toxicology section of the application.

• Identifying and controlling impurities in complex UPLC separations
• Orthogonal methods and detection
• The application and use of assays and purity determination of drug substances
• Non-denaturing methods and purity
  

• Do you see substantial differences in yield when using different template DNA sequences?
• Do you see substantial differences in dsRNA when using different template DNA sequences?
• Do you have purification processes that work as a platform, for different RNA sequences?
• Does your formulation/encapsulation process work as a platform, for different RNA sequences?
• Are there any best practices that you are willing to share?​

The solid-phase oligonucleotide synthesis based on sequential coupling of phosphoramidite monomers is a well-established industrial manufacturing process, currently performed routinely on kilo-scale mainly due to limitations of synthesis and purification processes. Novel approaches towards more efficient, scalable, and sustainable large-scale manufacture will be discussed supporting future commercialization of the expanding range of high-volume siRNA therapeutics.

End-to-End (aka, from-seed-to-unit-dose) manufacturing represents the next generation of biopharmaceutical manufacturing processes for both large and small molecules. It is recognized by regulatory authorities as a key emerging technology. The US FDA has approved various small molecule products and recently issued draft guidance for industry on continuous manufacturing.  The International Council for Harmonization issued (ICH) Q13 and is adopted by the European Medicines Agency (EMA) to support the technology and give guidance as well. Companies can add flexibility and maximize the value of process analytics ICH Q14 [6] to enhance E-2-E manufacturing.

With the recent explosion in the field of mRNA-based therapeutics, there is growing need for the standardization of mRNA and LNP critical quality attributes (CQAs) that can guide the development of assays needed for characterization, QC release, and stability studies. Several initial efforts toward providing this guidance have been made by industry SMEs, creating an initial standard that can be continuously improved through cooperation with the agencies, academia, and industry.